Microbiology Procedure
  Home  Link to us  Resources  Site map  Search  Language

Index >> Cellular Components >> Biosynthesis of RNA Transcription

Biosynthesis of RNA Transcription

Biosynthesis of RNA Transcription - It is now known that DNA itself is not the direct template that orders the amino acid sequences in proteins. Instead, the information in DNA is first transferred to RNA which then serves as a template for protein synthesis (See, breaking of the genetic code).

RNA, as mentioned earlier, is chemically very similar to DNA except for the presence of ribose and uracil instead of deoxyribose and thymine in DNA.
The synthesis of RNA occurs on DNA templates. The DNA strands at one or more stages separate and function as templates for RNA synthesis by attracting complementary base pairs.Direct evi­dence to support that RNA synthesis needs DNA templates came from the discovery of the enzyme RNA polymerase (DNA dependant RNA polymerase).

This enzyme has been found to exist in all cells and links together ribonucleotides by catalysing the formation of the internucleotide 3'-5' phosphodiester bonds that hold the RNA backbone together . This enzyme functions only in the presence of DNA and the RNA thus synthesized has a complemen­tary sequence to that of the DNA template.

DNA dependant RNA polymerase is a multisubunit complex. The active form consists of five different polypeptides, namely β,β’,α2,ω and δ with molecular weights of 150,000; 160,000; 90,000; 40,000; 10,000 respectively Each peptides except α, appears only once giving the complete holoenzyme β,β’,α2,ω and δ) with a molecular weight around 0.5 X 106 daltons.

The enzyme without the sigma unit (δ) is called the core enzyme and has the ability to catalyse RNA synthesis. The sigma unit has no catalytic function but has a role in recognition of start signals. In vitro experiments have shown that in the absence of sigma, the core enzyme initiates RNA synthesis randomly along both strands of the DNA However, when the sigma subunit is present, the correct strand is selected.

In the presence of the sigma subunit, the enzyme binds very strongly to the DNA regions which contain the start signals (promotors). These promotor regions apparently are not transcribed but only serve as RNA polymerasc binding sites. Much variation has been found in both the length and sequence of promotor regions. The binding sequences are however, rich in AT as is secn in a variety of bacteria and fungi. After chain elongation commences, the sigma subunit dissociates from the core enzyme-DNA-nascent-RNA complex and becomes free to attach another core molecule. RNA synthesis is terminated by the presence of stop signals which exist at specific points along the DNA template.

Two types of signals exist One, read by the RNA polymerase by itself and rich in A residues, the other recognized by the rho factor. These two together ensure that R NA chain termination occurs. The base composition of the enzymatically synthesized RNA has been compared with the base composition of the DNA templates and this has been found to be similar. It is also now known that, When double helical DNA serves as a template only one strand is copied. This has been directly verified using SP-8 bacteriophage of Bacillus subtilis. The two complement­ary DNA strands of this virus have different, base composition and, therefore can be easily separated by heating and centrifugation. Using this DNA in cell free systems along with RNA polymerase and nucleotides, the RNA synthesized has been tested for hybridization with either of the two strands. It has been found that DNA­-RNA hybrids are formed only with one strand of the DNA indicating that only one strand is copied during transcription. Each RNA chain, like the DNA chain has a direction defined by the orientation of the sugar phosphate backbone. It is now known that RNA chains also grow in the 5'-3' direction. Newly inserted nucleotides are found at the 3' end while the ppp group is found attached at the 5' end

 

Home | Site map | Submit Article | Resources | Search